Objective:To investigate molecular characterization and the immune responses of Manila clam hydramacin(Rp-hdmc).Methods:cDNA sequence of hydramacin was isolated from Manila clam transcriptome database.Molecular charac...Objective:To investigate molecular characterization and the immune responses of Manila clam hydramacin(Rp-hdmc).Methods:cDNA sequence of hydramacin was isolated from Manila clam transcriptome database.Molecular characterization of hydramacin cDNA was performed by BLAST and SWISS-MODEL bioinformatics programs.Tissue-specific expression and transcriptional regulation after Vibrio tapetis challenge was done by quantitative real time PCR.Results:Rp-hdmc has 291 bp open reading frame(ORF),encoding 97 amino acids with a mature hydramacin consisting of 77 amino acid residues.In un-challenged clam,Rp-hdmc was constitutively expressed in all tested tissues and the highest expression level was detected in gill.After pathogenic bacteria Vibrio tapetis challenge,Rp-hdmc mRNA was up-regulated in gill and hemocytes.Conclusions:We identified hydramacin cDNA(Rp-hdmc)from mollusk Manila clam that shows the characteristic features of hydramacin sequence.It has eight cysteine residues with four disulfide linkages,three helices and two β-strands in secondary structure.Expression results after V.tapetis challenges suggest that Rp-hdmc is involved in immune response against pathogenic bacteria.展开更多
基金Supported by the National Research Foundation(NRF)program of Ministry of Education,Science and Technology,Republic of Korea under research grant title of the Studies on the Manila clam(Ruditapes philippinarum)internal defense system(NRF-2011-0022671).
文摘Objective:To investigate molecular characterization and the immune responses of Manila clam hydramacin(Rp-hdmc).Methods:cDNA sequence of hydramacin was isolated from Manila clam transcriptome database.Molecular characterization of hydramacin cDNA was performed by BLAST and SWISS-MODEL bioinformatics programs.Tissue-specific expression and transcriptional regulation after Vibrio tapetis challenge was done by quantitative real time PCR.Results:Rp-hdmc has 291 bp open reading frame(ORF),encoding 97 amino acids with a mature hydramacin consisting of 77 amino acid residues.In un-challenged clam,Rp-hdmc was constitutively expressed in all tested tissues and the highest expression level was detected in gill.After pathogenic bacteria Vibrio tapetis challenge,Rp-hdmc mRNA was up-regulated in gill and hemocytes.Conclusions:We identified hydramacin cDNA(Rp-hdmc)from mollusk Manila clam that shows the characteristic features of hydramacin sequence.It has eight cysteine residues with four disulfide linkages,three helices and two β-strands in secondary structure.Expression results after V.tapetis challenges suggest that Rp-hdmc is involved in immune response against pathogenic bacteria.